Delivery system for biological component

ABSTRACT

A controlled release delivery system composition and method for oral administration of a biological component is disclosed. Preferably, a bacterium is delivered, and more preferably the bacterium is probiotic in nature.

CROSS-REFERENCE(S) TO RELATED APPLICATION(S)

This application claims the benefit of U.S. patent application Ser. No.10/261,639 entitled “Delivery System for Biological Component” filedSep. 30, 2002, which is hereby incorporated to this application by thisreference.

BACKGROUND

The present invention is directed to a controlled release solid dosageform for biological components. In addition, the invention is directedto a method of delivery of beneficial microorganisms over an extendedtimeframe.

As a substance passes through the human gastrointestinal (GI) tract itis subjected to a wide range of pH values ranging from the neutral pH ofthe mouth, to the acidic conditions of the stomach, to the 5.0-7.5 pHrange of the intestinal tract. Because the majority of biologicallyactive components are highly pH sensitive, these changes in pH can causesignificant effects upon the stability of the biological component andtheir ability to function in vivo. For example, many proteins denaturein acidic environments; once denatured, their biological activity, ifpresent, significantly differs from the non-denatured state. For abiological component (BC) to be functional, it must survive thegastrointestinal tract with minimal exposure to pH fluctuations.Further, BCs are also sensitive to enzymatic degradation. For example,one barrier to the oral administration of insulin is its susceptibilityto enzymatic degradation.

The oral administration of biological components without a controlledrelease system has as a significant disadvantage not allowing for thebiologic to by-pass the low pH and enzyme-rich environment of thestomach, thereby potentially decreasing the viability of the BC. Forthose devices which employ an enteric coating mechanism to survive thegastric environment, the shortcomings may be two-fold. First, theprocess of coating the dosage form or its contents may result insignificantly lowered viability of the BC. Second, the downfall ofmerely by-passing the stomach is the explosive delivery of the biologicimmediately upon exiting the stomach. This non-specific delivery isineffectual and primitive in view of certain delivery needs ofbiological components because the bioavailability of BCs is often sitedependent. Biological components may be targeted either throughmodification of the biologic itself or through the controlled release ofthe biologic within a desired physiologic window. One such biologicalcomponent that displays such site-specificity is the lactic acidbacteria, Lactobacillus Acidophilus (a probiotic). L. Acidophilus is oneexample of other probiotics, including Lactobacillus bulgaricus,Lactobacillus casei subsp. Rhamnosus, Lactobacillus casei subsp. Casei,Lactobacillus salivarius, Lactobacillus brevis, Lactobacillus reuteri,Lactococcus lactis subsp. Lactis, Enterococcus faecium, Lactobacillusplantarum, Streptococcus thermophilus, Bifidobacterium infantis,Bifidobacterium Bifidum, Bifidobacterium longum, Saccharomycesboulardii, and various modified soil organisms.

Each strain of L. Acidophilus will attach at a different location of theintestinal tract, preferentially attaching within a region eitherslightly proximal or distal to other L. Acidophilus strains. Thesepreferential regions of attachment are of particular importance relativeto employing the bacteria as delivery systems for genomic or proteomictherapy, whether directly or as carriers for other vectors containinggenetic or proteomic biologicals.

Beneficial microorganisms, for example, but not limited to,gastrointestinal flora such as lactic acid bacteria and yeast are anessential constituent of metabolism and immune response. Supplementationof beneficial microorganisms is a valid mechanism for replacement offlora lost due to antibiotic treatment, enhancement ofnaturally-occurring levels of beneficial flora, enhancing competitiveinhibition and otherwise preventing enteropathogens, and altering themetabolism of ingested substances. Probiotics are one example ofbeneficial microorganisms.

Solid oral dosage forms employing controlled release have beenincreasingly demonstrated to be beneficial to the administration ofpharmaceutical compounds, enhancing safety and consumer compliance,minimizing side effects and providing new therapeutic benefits. The fourgeneralized platforms for controlled release solid oral dosage formsare. diffusion, reservoir, pore-forming wax, or coated-bead systems. Fewhave been applied to BCs due to high development costs, bioavailabilityissues, and stability of the dosage BC within the dosage form. In thepast, enteric coating technologies and other mechanisms of delayedrelease have been limited to features with explosive delivery after thestomach.

Controlled release delivery systems can take many forms includingpolymeric matrix systems, wax matrix systems, multi-particulate systems,and combinations thereof. The most commonly used delivery systems can bebroadly classified as diffusion, reservoir, pore-forming wax, orcoated-bead systems. Diffusion devices are composed of a drug dispensedin a polymer which diffuses from the entire physical tablet. Reservoirdevices usually consists of a semi-permeable barrier which is involvedin the release of the active from a core site within the tablet.Coated-bead systems employ an enteric or pH-sensitive coating ofaggregated particles of the active ingredient packaged in capsule form.Pore-forming wax systems incorporate the active ingredient into a waxbase and rely upon the rate of diffusion to control the release of theactive ingredient.

In tableted, pore forming wax matrices, the BC and a water-solublepolymer are introduced into a wax or wax-like compound such as paraffinor guar gum, and then placed in an aqueous environment so as to allowthe water-soluble polymer to dissolve out of the wax, resulting in theformation of pores. Upon contact with the gastrointestinal fluid, thepores facilitate diffusion-mediated release of the BC. The rate ofrelease of the BC is dependent upon non-linear erosion.

Coated-bead systems are one of the few delivery systems available inboth tablet and capsule form. The BC encased within a bead using one ofthe variety of processes available, such as spheronization-extrusion orcoating of non-pareils. The coated-BC is then further coated with anenteric coating or employed in a blend of coated-beads with differingrelease rates for extended release formulations. The BC may also beblended or granulated with polymers before coating to provide anadditional level of control. The coated-beads themselves may also becombined with polymers to create a hybrid diffusion or wax-based system.Coated-bead systems are complex to manufacture, requiring large numbersof excipients, use of solvents, and long manufacturing time. The use ofsuch solvents and the manufacturing processes required to apply suchsolvents may expose the BC to adverse environmental conditions and causea loss of the viability of the BC. This is especially concerning in thecase of lyophilized BCs, where any exposure to moisture may causesignificant decreases in viability.

An example of a reservoir system is the push-pull osmotic pump. Theseosmotically-controlled delivery systems feature a bi-layer tablet coatedwith a semi-permeable membrane possessing a laser-bored orifice throughwhich the BC is pushed as aqueous solution is absorbed into the tablet.There are a number of osmotic delivery systems on the market that workvia a similar physical principle; these osmotic systems produce veryreplicable, linear release. Manufacturing this system is definitivelynon-conventional, requiring specialized equipment and additionalprocessing steps. The inherent complexity of the design adds acorresponding complexity to the development and scale-up of any osmoticmembrane product.

The diffusion tablet systems rely on hydrophilic polymer swelling forcontrol of BC release. Polymer systems can be sub-classified asconventional hydrogel systems and modified polymer systems. Conventionalhydrogel systems rely upon the penetration of water to form of agel-like phase through which the bioactive agent is released. Thesesystems often incorporate the BC in a single polymer such aspolyethylene oxide or hydroxypropyl methylcellulose. In the case ofmodified polymer systems, polymers with differing physicalcharacteristics—such as one that is hydrophilic (e.g. HPMC), and onethat is pH-dependent in its swelling characteristics, (e.g. pectin)—arecombined with the BC. When these polymers interact with dissolutionmedia, a transition phase or interfacial front develops, forming agradually dissociating semi-solid core surrounded by a gel peripherythat allows the BC to be increasingly released as the matrix hydrates.The movement of the dosage form through the gastrointestinal tract,through regions of increasing pH, permits further swelling and erosionof the matrix, culminating in complete release of the BC and completedissolution of the dosage form.

Prior art formulations cannot deliver beneficial microorganisms over anextended time period or to targeted individual regions of GI tract.Prior art formulations require coating processes to achieve gastricbypass. Further, prior art formulations fail to provide mechanisms forpH control thereby rendering pH sensitive strains much less viable dueto variations in GI pH. Further, prior art formulations lack mechanismsof isolating the BC from enzymatic degradation. Prior art formulationslack mechanisms to increase the stability of the dosage form itselfthrough water sequestration of available water. Prior art formulationsutilizing dietary fiber as a carrier require too large a volume forefficient oral dosage form manufacture. These and other limitations andproblems of the past are solved by the present invention.

SUMMARY

The present invention provides controlled release delivery systems fororal administration of a biological component. Further, a beneficialmicroorganism is delivered; the probiotic being bacterial in nature.

One embodiment of a controlled delivery system includes a hydrogel ormodified matrix formed from an excipient of one or more hydrophilicpolymers, polysaccharides, galactomannan gums, resins, polyethylenederivatives or hydrolyzed proteins, either alone or in combination, inwhich is disposed biological components, in one aspect beneficialmicroorganisms, and in yet another aspect lyophilized bacteria and theirassociated lyophilized carrier proteins. Optionally, the delivery systemincludes one or more additional release modifying excipients from thesame group of hydrophilic agents for the purpose of attenuating therelease of the lyophilized ingredients with pH-specific orenzyme-specific agents, and optionally, one or more physiologicallyacceptable electrolytic substances included for the purpose of pHcontrol or available water-sequestration.

In another embodiment, the controlled delivery system includes a waxmatrix composed of one or more inert insoluble waxes, polymers and/orfillers, alone or in combination, in which is disposed pore formingexcipients and the beneficial microorganisms, in one aspect inlyophilized form and their associated lyophilized carrier proteins.

In yet another embodiment of a controlled delivery system includes amulti-particulate system in which a plurality of granules, coated beadsor coated non-pareils are distributed within the dosage form in either asimple or an modified polymer matrix or for the purposes of controlledrelease of beneficial microorganisms, in one aspect in lyophilized formand their associated lyophilized carrier proteins.

In another embodiment, a process for making an extended release dosageform, such as a tablet or capsule, from a pre-blend including mixing abeneficial microorganism with one or more polymers, gums, resins,polyethylene derivatives, or hydrolyzed proteins for the purpose ofcontrolled release; the optional addition of physiologically acceptableelectrolytic substances for the purpose of regulating pH within thedosage form; and the optional inclusion of available water-sequesteringelectrolytic species for the purpose of increasing the stability of thedosage form itself.

In another embodiment of the method of making an extended release dosageform, such a tablet or capsule, includes mixing a beneficialmicroorganism with a pre-blend of one or more controlling excipients,fillers, desiccants, and flow agents that has been mechanically,chemically, or otherwise dried to reduce the available water present forthe purpose of preventing undesirable interactions of the beneficialorganisms and hydrophilic agents with any available water within thedosage form.

The system generally includes a hydrophilic agent, an electrolyte, and abiological component (BC), and may optionally include fillers, releasemodifying agents, desiccants, and flow agents.

In one embodiment, a delivery system for disclosed including ahydrophilic or hydrophobic agent and the BC.

In another embodiment, a delivery system is disclosed including ahydrophilic agent, an electrolytic agent, and the BC.

In yet another embodiment, a delivery system is disclosed including ahydrophilic agent, a release modifying agent, and the BC.

In yet a further embodiment, a delivery system is disclosed including ahydrophobic agent, a release-modifying agent, and a BC.

In yet a further embodiment, a delivery system is disclosed including ahydrophilic agent, a release-modifying agent, and a BC.

In yet a further embodiment, a delivery system is disclosed including ahydrophilic agent, electrolyte, and BC.

In yet a further embodiment, a delivery system is disclosed including ahydrophobic agent, release-modifying agent, electrolyte, and BC.

In yet a further embodiment, a delivery system is disclosed including ahydrophilic agent, release-modifying agent, electrolyte, and BC.

The controlled release formulations for beneficial microorganisms havemany advantages over the current art. Targeted delivery of beneficialmicroorganisms, such as probiotic bacteria, allows for dispersion ofprobiotic organisms within regions of optimal attachment that may bespecific to a given strain or therapeutic goal. One advantage isachieving gastric bypass for the biological contents. Another advantageof the system disclosed is the maintenance of a constant pH within thedosage form surrounding the beneficial microorganisms, allowing anoptimal microenvironment for reconstitution of lyophilized ingredientsto be created, thereby maximizing viability of the lyophilizedingredients released into the GI tract. Another advantage of the systemdisclosed is the inclusion of available water-sequestering electrolyticspecies, an optimal microenvironment may be maintained during storage,thereby increasing the stability of the dosage form itself. Furtheradvantages of the system disclosed are it requires only dry blend anddirect compression steps, the system is easily transferable to sites ofmanufacture and relies on only conventional tableting or encapsulationequipment for production. Because this system is relatively independentof the biological components employed in formulation, targeted deliveryof genetically modified bacteria or other beneficial microorganisms isalso possible.

One advantage of the present system is the controlled release of thebacteria from the dosage form into the surrounding environment. Anotheradvantage of the present system is the maintenance of a constant pHwithin the dosage form itself through the use of physiologicallyacceptable electrolytic substances.

Yet another advantage of the present system is the controlled exposureof the bacteria within the dosage form to aqueous media throughcontrolling the hydration rate of the dosage form via polymerdisentanglement.

Yet another advantage of the present system is an increase in thestability of the dosage form and the viability of the contents throughthe inclusion of available water-sequestering electrolytic species.

Yet another advantage of the present system is its manufacturability: adry-blend and direct compression form of tablet manufacture and adry-blend and direct fill form of capsule manufacture. Most advantageousis the absence of any processes which introduce moisture (such ascoating or granulation) that may decrease the in vivo viability of thebiological component.

The invention will best be understood by reference to the followingdetailed description of the preferred embodiment. The discussion belowis descriptive, illustrative and exemplary and is not to be taken aslimiting the scope defined by any appended claims.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows the effects of hydrophilic agents on the controlled releaseof viable beneficial microorganisms into the small intestine frommonolithic tablets.

FIG. 2 shows the effects of the addition of electrolytes on thecontrolled release of viable beneficial microorganisms into the smallintestine from monolithic tablets.

FIG. 3 shows the effects of the addition of pH- and enzyme-sensitiveagents on the controlled release of viable beneficial microorganismsinto the small intestine from monolithic tablets.

FIG. 4 shows the effects of the addition of pH- and enzyme-sensitiveagents on the controlled release of viable beneficial microorganismsinto the small intestine from monolithic tablets.

FIG. 5 shows the effects of electrolytes and pH- and enzyme-sensitiveagents on the controlled release of viable beneficial microorganismsinto the small intestine from monolithic tablets.

FIG. 6 shows the capacity for the controlled release of viablebeneficial microorganisms over extended durations from monolithictablets.

FIG. 7 shows the controlled release of beneficial microorganisms over anextended duration of 8 hours from monolithic tablets.

FIG. 8 shows the controlled release of beneficial microorganismsspecific to the lower intestinal tract over an extended duration of 12hours from monolithic tablets.

FIG. 9 shows the effects of a hydrophilic matrix on the controlledrelease of viable beneficial microorganisms into the small intestinefrom capsules.

FIG. 10 shows the capacity for geometric scalability and tablet size andshape variation in the present invention and the effect of such changeson the controlled release of viable beneficial microorganisms into thesmall intestine from monolithic tablets.

FIG. 11 shows the drying the excipients prior to tableting and theeffect of such changes on the controlled release of viable beneficialmicroorganisms into the small intestine from monolithic tablets.

FIG. 12 shows the effects of a hydrophilic matrix employing hydrophilicpolymers of differing viscosities on the controlled release of viablebeneficial microorganisms into the small intestine from capsules.

FIG. 13 shows the effects of physiologically acceptable electrolyticsubstances on the stability of the dosage form.

DETAILED DESCRIPTION

A delivery system is disclosed for the controlled release of abiological component into the surrounding environment. Controlledrelease delivery systems include those systems capable of site specificdelivery, extended release, sustained release, delayed release, repeataction, prolonged release, bimodal release, pulsitile release, modifieddelivery, pH sensitive delivery, and/or target specific delivery, amongothers. The biological components include, but are not limited to,beneficial microorganisms, such as probiotic bacteria. The solid dosageform may take the form of a tablet, capsule, wafer, or sachet, and isnot limited to, an orally administered dosage form such as a tablet orcapsule.

As used herein, a delivery vehicle, for example a homogenouslydistributed matrix, is made up of hydrophilic agents and/or ahydrophobic agents. Hydrophilic agents include swelling, viscosityincreasing, gel strength enhancing agents. Hydrophobic agents includewaxes and other inert materials, such as ethylcellulose or carnauba wax.More particularly, the hydrophilic agent is selected from at least oneof the group, but not limited to: a) a starch selected from the groupconsisting of corn, rice, or potato starch; b) a hydrophilic gum,polysaccharide or galactomannan selected from the group consisting ofpectin, agar, dextran, carageenan, tragacanth gum, locust beam gum,acacia gum, guar gum, xanthan gum, ghatti gum, alginic acid or sodiumalginate; c) a cellulose derivative selected from the group consistingof methylcellulose, carboxymethylcellulose, sodium starch glycollate,sodium or calcium carboxymethylcellulose, hydroxyethyl methylcellulose,hydroxypropyl methylcellulose, ethylhydroxy ethylcellulose,ethylmethylcellulose, hydroxyethylcellulose, cellulose acetate phthalateor microcrystalline cellulose; d) silica, aluminum silicate, magnesiumsilicate, aluminum magnesium silicate, sodium silicate or feldspar, e)aluminum hydroxide; f) a protein selected from the group consisting ofgelatin or casein; and g) a polymer selected from the group consistingof acrylate, carboxypolymethylene, a polyalkylene glycol orpolyvinylpyrrolidone. In one aspect, the hydrophilic polymers areselected from the group of cellulose derivatives such asmicrocrystalline cellulose (MCC), hydroxypropyl methylcellulose (HPMC),or hydroxypropyl cellulose (HPC), or from gums and polysaccharides suchas guar gum or maltodextrin.

As used herein, optionally, the system may include agents added to aidin gastric bypass or modify the release profile of the BC due topH-specific swelling characteristics or site-specific enzyme degradationwithin the GI tract. These agents may include but are not limited to atleast one of alginate, polysaccharides such as such as gelatin orcollagen, guar gum, xanthan gum, pectin, heterogeneous protein mixtures,and polypeptides. The polysaccharides may be pectin and/or an alginatesalt, among others. The galactomannan gums may be guar gum, xanthan gumand/or locust bean gum, among others. The polyethylene derivatives maybe polyethylene oxide (PEO) and/or polyethylene glycol (PEG), amongothers. The hydrolyzed proteins may be gelatin and/or collagen, amongothers.

As used herein, BC includes agents such as microbes, DNA, RNA, protein,modified soil organisms (organisms that compete with lactic acidbacteria) bacteria, and biopharmaceuticals. The biological component maybe viable or non-viable. The BC may be a beneficial microorganism (orprobiotic); and yet in another aspect, the beneficial microorganism isbacterial in nature. The term “probiotic” refers to ingestedmicroorganisms that can live in a host and contribute positively to thehost's health and well being.

As used herein, the electrolytes may be at least one of sodium,potassium, or calcium salts, among others. Through the inclusion ofphysiologically acceptable electrolytes, the buffered environment allowsreconstitution and release to occur under optimal pH conditions forbacterial viability. The interaction between electrolytes and ahydrophilic agent may allow not only the pH-independent release of theBC, but also allows for the internal pH of the dosage form to remainconstant. It is this constant internal pH that contributes significantlyto the stability of the biological contents in-vivo.

Optionally, physiologically acceptable salts may be introduced to thebacterial freeze-dried product (FDP) during lyophilization at a ratio of1.0:0.1 to 1.0:25 bacterial FDP to salt. The system ensures themaintenance of a constant pH within the dosage form itself and acts as acryoprotectant during the freeze-drying process to prevent lysing of thecell.

As used herein, the system may optionally include a desiccant. Thedesiccant may include, but is not limited to, sodiumcarboxymethylcellulose, calcium carboxymethylcellulose, colloidal silicadioxide, and combinations thereof. The disintegration agent may include,but is not limited to, croscarmellose sodium sold as Solutab™ availablefrom Blanver Farmoquimica LTDA and crosprovidone (insolublepolyvinylpyrrolidone) sold as Kollidon CL™ available from BASF.

As used herein, the system may optionally include flow and tubingagents. The flow agents may include, but are not limited to, magnesiumstearate and stearic acid.

In a first embodiment, the delivery system includes a swellinghydrophilic agent and a BC. It is based on the homologous distributionof the various components within a solid matrix dosage form. The systemallows for a controlled exposure of the BC within the dosage form to anaqueous media by controlling the hydration rate of the dosage form viapolymer disentanglement and matrix erosion. Optionally, the system mayalso include a physiologically acceptable electrolyte, a releasemodifying excipient such as a gum or polysaccharide, a desiccant, andflow or tubing agents, alone or in combination. Electrolytes can providea mechanism for available water-sequestration to increased stability ofthe dosage form and the viability of its contents. Desiccants may alsobe used to sequester available water for a similar purpose. Releasemodifying excipients, such as gums and polysaccharides, may be used toinduce site-specific release through pH-specific swelling orsite-specific enzymatic degradation. Flow or tubing agents may be usedto improve the manufacturability. This may also result in decreased lossof viability during manufacture due to compression and heat resultingfrom powder flow, tableting, and encapsulation.

In one aspect of the embodiment, the BC may be a probiotic pre-blend,which can be blended with a carrier. The carrier may be, but is notlimited to, monosaccharides or polysaccharides, such as maltodextrin,swellable polymers, such as hydroxypropyl methylcellulose, inertfillers, such as microcrystalline cellulose or di-calcium phosphate, orother inert substances, such as carnauba wax. In the aspect wherein acarrier is included, the carrier may function to assist in thecontrolled release of the BC, to aid in the manufacturability of thedosage form, or to increase the stability of the dosage form.

The delivery system can be a readily manufacturable solid dosage form.In one aspect, the dosage form is in the form of a monolithic tablet orcapsule. When a tablet or capsule, it may be administered orally,anally, and vaginally, among other routes. In one aspect, the dosageform is a monolithic tablet created from a direct-compressible dry blendwhich does not require processes, such as enteric coating, granulation,or spray drying, that expose the BC to temperatures that might cause anybiological contents to be damaged. However, provided such coating orgranulation processes are carried out in a manner that do not damage thebiological contents, nor adversely affect the hydration state of thematrix, they may be amenable.

Release of the biologic into the surrounding environment may beaccomplished through a rate-controlled hydration and subsequent swellingof hydrophilic agents. The release of the biologic is determined by theerosion rate and polymeric disentanglement of the swollen hydrophilicmatrix. Without subscribing to a particular theory of kinetics, theswelling of the hydrophilic matrix is retarded by a plurality of layersof viscous gelled hydrophilic agents; these gel-states result from theinteraction of the hydrophilic agents with the penetratinggastrointestinal fluid. While primarily erosion dependent, the gradualhydration and gelling reaction within the hydrophilic matrix allows fora highly reproducible, programmable release pattern. The programmabilityof the system allows for nearly any physiologically relevant releasepattern to be accomplished. Mathematical treatment of the hydrophilicmatrix swelling, erosion, and ensuing release of BC can be determined,though each model will be representative of the particular componentsspecific to each formulation. This can be accomplished without the needfor undue experimentation. Formulation specific to the physicalcharacteristics of each BC and the desired release profile can beaccomplished through both theoretical and empirical means, allowingdissolution of the system and BC release to occur in a specificphysiologic region. Release of contents in a given region of the GItract is accomplished by the slowly hydrating hydrophilic matrixcontaining the biological actives segregated from the externalenvironment until the desired physiologic region of release, which maybe employed to achieve gastric bypass. Consideration of both the areaand duration of release is essential in formulation so as to program thesystem with an appropriate ratio of components to ensure the desiredrelease profile.

The homologous distribution of BCs within the hydrophilic matrixprovides protection from the fluctuations in pH and exposure toenzymatic degradation present in external environment. When lyophilizedmicroorganisms are delivered, this isolation from the outsideenvironment allows the bacteria to remain in lyophilized stasissignificantly longer than with conventional immediate release dosageforms.

In another embodiment, when physiologically acceptable electrolytes areincluded into the delivery system, the electrolyte maintains anintra-dosage form pH irrespective of the external pH. This internal pHmay be modified through the selection of electrolytes that are bothphysiologically-acceptable for human consumption andphysiologically-appropriate to individual BCs. When deliveringlyophilized beneficial microorganisms, this internal pH may be selectedto create an optimal environment for the reconstitution of thelyophilized organisms. Such an environment may result in an increase inviability during the reconstitution process, and moreover, may limit theexposure of the lyophilized microorganism to fluctuations ingastrointestinal pH, resulting in an increase in organism viabilitywhile the matrix is in a hydrated state and prior to the organismsrelease into the environment.

The addition of physiologically-acceptable electrolytes may also beemployed to aid in available water-sequestration. When deliveringlyophilized beneficial microorganisms, this is especially useful, asinteractions with any available water—such as the available waterpresent in the constituent controlling excipients, flow agents anddesiccants—may result in inadvertent, premature reconstitution prior torelease in the gastrointestinal environment. Premature reconstitutionfrom a lyophilized state causes the organisms to begin metabolizingavailable sources of energy; the constituents of the delivery systemprovide very limited sources of energy and when these locally availablesources of energy are exhausted, the organisms expire. The metabolicbyproducts of prematurely reanimated organisms may also have a negativeimpact on the viability of the remaining, non-reanimated organisms. Whendisposed in a homogeneous manner throughout the dosage form,electrolytic substances that have a higher degree of hydrophilicity thanthe other constituents of the delivery system surrounding them maypreferentially hydrate, decreasing or preventing the re-hydration of thelyophilized agents. An example of a system not including an electrolyteis a system that is dependent upon erosion as its release mechanism, orone in which the maintenance of a constant pH within the dosage form isnot desired; lyophilized beneficial microorganisms and hydrophilicagents do not require an electrolyte to make a controlled release dosageform capsule. Another example that does not require an electrolyte iswhere the controlled release of non-viable beneficial microorganisms(such as non-viable bacterial biomass) is sought as the primary functionof the dosage form.

In another embodiment of the delivery system, the addition of releasemodifying excipients, such as hydrophilic polymers or gums demonstratingpH or enzyme sensitivity, may be employed to alter the swelling orerosion characteristics of the matrix, such as the initiation ofswelling or the rate of erosion of the matrix. These release modifyingexcipients function in combination with the hydrophilic agent to controlthe release of the biological component. These excipients may beemployed to reduce the amount of exposure to the gastric environment byreducing matrix swelling during exposure to gastric pH or during thetime the dosage form is expected to transit through the stomach andpylorus. These release modifying excipients may be selected for their invivo degradation characteristics that occur in localized regions of thegastrointestinal tract. The release modifying agent, when used alone,may function as the hydrophilic agent. One example of this, among many,is that pectin mainly breaks down at the higher pH and enzyme richenvironment of the large intestine, thus it can be employed alone as thehydrophilic agent if a greater proportion of lower intestinal tractdelivery was desired. Another example among others it that gelatinlargely breaks down in the small intestine. With regards topharmaceutical controlled release formulations, the location of polymerbreakdown is of special significance as bioavailability is determined bythe amount of drug released within a given timeframe relative to aphysiological site of absorption specific to that type of compound. Thedelivery of biological components is essentially similar in intent,given localized sites for absorption and adsorption. When deliveringbeneficial microorganisms, the inclusion of release modifying excipientswhose swelling characteristics are pH dependent, specifically compoundsthat preferentially swell in environments above pH 1.0-1.5, is usefulfor the delivery of lactic acid bacteria that are susceptible toviability losses when exposed to low pH. The low-pH environment willinhibit swelling, thus retarding both beneficial microorganism releaseand acid-penetration into the dosage form. The inclusion of releasemodifying excipients whose erosion is enzyme-dependent, specificallycompounds that degrade preferentially in the presence of lowerintestinal tract enzymes, is useful for the delivery of lactic acidbacteria whose attachment site is distal to the location of the enzymes.

In another embodiment of the delivery system, the system is apore-forming wax matrix composed of one or more inert insoluble waxes,polymers or fillers in which is disposed pore forming excipients and theactive lyophilized bacteria and their associated lyophilized carrierproteins. Hydrophilic agents may be included with hydrophobic agents tomake pore forming wax matrices.

In yet another embodiment, the system may include a multi-particulateplurality of granules, coated beads or coated non-pareils aredistributed within the dosage form in either an active polymer matrix orimmediate release matrix for the purposes of controlled release of thelyophilized active ingredients.

In one embodiment, the dosage form disclosed is formed from a pre-blend.When a monolithic tablet, the pre-blend is mixed using dry-blendtechniques known to those skilled in the art, and the dosage form iscreated using a direct compression process. Employing a pre-blend thatis formed using dry-blend techniques is a significant improvement overthe use of blends resulting from granulation, spheronization-extrusion,or other processes that might expose the biological components tomoisture or solvents and potentially lower the viability of thebiological components. Employing a pre-blend that is capable of forminga monolithic dosage form using only the techniques ofdirect-compression, in the case of a tablet, or high speedencapsulation, in the case of a capsule, is a significant improvementover manufacturing processes that require multi-stage compression,multiple geometrically-altered components or coatings that might exposethe biological component to hazardous environmental conditions such assolvents, high forces of compression, excessive heat or undue physicalstress. When delivering lyophilized beneficial microorganisms,preventing the premature reconstitution of the organisms is important tomaintaining the in vivo viability of the organisms.

The dosage form disclosed may be formed from a pre-blend in which alyophilized biological component, for example a lyophilized beneficialmicroorganism, is mixed with a pre-blend of one or more controllingexcipients, fillers, desiccants, and flow agents that has beenmechanically, chemically, or otherwise dried to reduce the availablewater present for the purpose of preventing undesirable interactions ofthe beneficial organisms and hydrophilic agents with any available waterwithin the dosage form. The minimization of available water within thedosage form is intended to prevent unintentional or pre-maturereconstitution of the lyophilized organisms. The use of a pre-blend inwhich the non-lyophilized components are dried and subsequently blendedwith the lyophilized components, while not necessary for the creation ofa controlled release dosage form, is a significant improvement over theuse of either non-dried excipients that may contain enough availablewater to induce pre-mature reconstitution prior to in vivo release, orthe drying of a pre-blend containing both lyophilized andnon-lyophilized components, which exposes the lyophilized components toundue heat and may extensively reduce their in vivo viability.

Unless otherwise noted, all of the following embodiments are formulatedthrough standard dry blend and directly compression with an appropriatelubricant such as magnesium stearate or stearic acid.

In the first embodiment, a formulation is disclosed combining thebacterial lyophilized (freeze-dried) powder pre-blend (FDP) with asuitable hydrophilic agent such as HPMC, MCC, or PEO, in a ratio ofabout 1.0:0.1 to 1:25 FDP to hydrophilic agent.

In the second embodiment, a formulation is disclosed including bacterialFDP, hydrophilic agent, and a physiologically acceptable electrolytesuch as NaHCO₃, Na₂ CO₃, or Ca CO₃, in a ratio of about 1.0:0.1:0.1 to1:25:25 FDP to hydrophilic agent to electrolyte.

The third embodiment, a formulation is disclosed including bacterialFDP, a hydrophilic agent, and a release modifying agent in the form of ahydrophilic polysaccharide such as pectin, sodium alginate alginic acid,or a gum such as xanthan gum, guar gum, locust bean gum, or tragacanthgum, in a ratio of about 1.0:0.1:0.1 to 1:25:25 FDP to hydrophilic agentto polysaccharide or gum.

The fourth embodiment, a formulation is disclosed including bacterialFDP, a hydrophilic agent, a release modifying agent in the form of ahydrophilic polysaccharide or gum, and a physiologically acceptable saltin a ratio of about 1.0:0.1:0.1:0.1 to 1:25:25:25 FDP to hydrophilicagent to polysaccharide or gum to electrolyte.

The fifth embodiment, a formulation is disclosed including bacterialFDP, a hydrophilic agent, a release modifying agent in the form of ahydrophilic polysaccharide or gum, a physiologically acceptable salt,and an inert filler in a ratio of about 1.0:0.1:0.1:0.1:0.1 to1:25:25:25:25 FDP to hydrophilic agent to polysaccharide or gum toelectrolyte to inert filler.

In the sixth embodiment, a formulation is disclosed combining thelyophilized lactic acid bacteria pre-blend with a suitable hydrophobicagent such as carnauba wax, in a ratio of about 1.0:0.1 to 1:25 FDP tohydrophobic agent.

In the seventh embodiment, a formulation is disclosed includingbacterial FDP, a hydrophobic agent, and a physiologically acceptableelectrolyte such as NaHCO₃, Nat CO₃, or Ca CO₃, in a ratio of about1.0:0.1:0.1 to 1:25:25 FDP to hydrophobic agent to electrolyte.

The eighth embodiment, a formulation is disclosed including bacterialFDP, a hydrophobic agent, a physiologically acceptable electrolyte suchas NaHCO₃, Na2 CO₃, or Ca CO₃, and a release modifying agent in the formof a hydrophilic polysaccharide such as pectin, sodium alginate alginicacid, or a gum such as xanthan gum, guar gum, locust bean gum, ortragacanth gum, in a ratio of about 1.0:0.1:0.1:0.1 to 1:25:25:25 FDP tohydrophobic agent to polysaccharide or gum to electrolyte.

The dosage forms may be monolithic tablets or gelatin or vegetablecapsules for oral, anal, or vaginal delivery.

Methods

The formulations described below have been prepared in accordance withthe following methods. In these formulations, tablets were preparedusing a method of dry blending and direct compression using a Carverhydraulic press or a rotary tablet press. Evaluations were performedusing a USP Type II (paddle) dissolution apparatus.

Examples 1-5, 9, 10, and 12 were conducted by exposing the dosages to1000 mL 0.1N HCl for 2 hours at 50 RPM. The dosages were then removedand placed into peptone buffer medium and stomached, (the dosage form iscrushed and homogenized within the buffer media for the purpose ofenumerating the remaining bacteria in the tablet), after which a samplewas taken from the dissolution media. The samples were then plated onMRS and RCM media to discern viable colony forming units (CFU).

Example 6 was performed by exposing the dosages to 1000 mL USP HCl for 2hours at 50 RPM. The dosages were removed and placed into KH₂PO₄ bufferdissolution medium and the dissolution media was sampled at regularintervals. The samples were then plated on MRS and RCM media to discernviable colony forming units (CFU).

Examples 7 and 8 were performed by exposing the dosages to 1000 mL USPHCl (Ex. 7) or 0.1N HCl (Ex. 8) for 2 hours at 50 RPM. The dosage formswere removed and placed into KH2PO4 (Ex. 7) or peptone (Ex. 8) buffermedium and the dissolution media was sampled at regular intervals. Thesamples were then filtered, reacted with 4′,6-diamidino-2-phenylindole,and enumerated under UV-light.

Example 11 was performed using tablets produced from excipientsdesiccated in a fluid bed drier, mixed with the lactic acid bacteriapre-blend and flow agents, and tableted. The dosage forms were thenexposed to 1000 mL 0.1N HCl for 2 hours at 50 RPM. The dosages wereremoved and placed into peptone buffer medium and stomached, after whichthe dissolution media was sampled. The samples were then plated on MRSand RCM media to discern viable colony forming units (CFU).

Example 13 was performed using dosages packaged in foil sachets andexposed to ambient environmental conditions (25 degrees C., 60% RelativeHumidity) for 4 months and subsequently tested. The samples were removedto peptone buffer solution, stomached, and plated on MRS and RCM mediato discern viable colony forming units (CFU).

EXAMPLE 1

A monolithic tablet of approximately 382 mg having a hydrophilic agentand biological component (BC) was prepared as shown in Table 1, with thegroup A1 as the control group. In this example, the beneficialmicroorganism is the lactic acid bacteria pre-blend of lyophilizedpowder and starch, and the hydrophilic agent employed ismicrocrystalline cellulose (MCC), maltodextrin, hydroxypropylmethylcellulose (HPMC), or polyethylene oxide (PEO). The addition of thehydrophilic agent will retard the release of the BC from the dosageform. Stearic acid is included as a flow agent and silica is employed asflow agent and desiccant.

As shown in FIG. 1, the results of this example reflect a level ofcontrolled release granted through the use of a matrix comprised of ahydrophilic agent and a lyophilized BC. This controlled release is shownthrough a much higher level of viable lactic acid bacteria colonyforming units (CFU) delivered after exposure to gastric media than thecontrol. The use of less swellable hydrophilic agents such as MCC andmaltodextrin are associated with sufficient, but lower levels ofcontrol. A superior level of control is demonstrated in bothpolyethylene oxide and HPMC matrices. Thus, the hydrophilic agent is notlimited to a particular type of hydrophilic agent, so long as sufficientmatrix viscosity is achieved. TABLE 1 Al Dosage Formulas (mg) (CTRL) A2A3 A4 A5 Lactic acid bacteria pre-blend 150 150 150 150 150 HPMC 0 0 0200 0 PEO 0 0 0 0 200 MCC 0 200 0 0 0 Maltodextrin 0 0 200 0 0 StearicAcid 16 16 16 16 16 Silica 16 16 16 16 16 TOTAL WEIGHT 182 382 382 382382

EXAMPLE 2

A monolithic tablet of approximately 382 mg containing a hydrophilicagent, an electrolytic agent, and a biological component was prepared asset forth in Table 2, with B1 as the control group. The formulationemploys HPMC as the hydrophilic agent, NaHCO₃, Na₂CO₃ or NaH₂PO₄ as theelectrolytic agent, and the lactic acid bacteria pre-blend oflyophilized powder and starch as the biological component (BC). Theaddition of NaHCO₃, Na₂CO₃ or NaH₂PO₄ establishes the pH within thedosage form. Stearic acid is included as a flow agent and silica isemployed as flow agent and desiccant.

This example demonstrates, as shown in FIG. 2, that the internal pH ofthe dosage form is altered by the presence of an electrolyte, affectingthe amount of viable CFU delivered. This establishment of a particularinternal pH is associated with differing levels of viability for a givenreconstituted lyophilized organism. In particular, formulation B2, whichcontains Na₂CO₃, provides an internal pH which aides in thereconstitution of viable lactic acid bacteria. TABLE 2 Dosage Formulas(mg) B1 (ctrl) B2 B3 B4 Lactic acid bacteria pre-blend 150 150 150 150HPMC 0 100 100 100 PEO 0 0 0 0 NaHCO₃ 0 100 0 0 NaHCO₃ 0 0 100 0 NaH₂PO₄0 0 0 100 Stearic Acid 16 16 16 16 Silica 16 16 16 16 TOTAL WEIGHT 182382 382 382

EXAMPLE 3

A monolithic tablet of approximately 382 mg containing a hydrophilicagent, a release-modifying excipient, and a BC was prepared as shown inTable 3, with C1 as the control group. The hydrophilic agent employed isHPMC, the release-modifying excipient employed is pectin or gelatin, andthe lactic acid bacteria pre-blend of lyophilized powder and starch isthe BC. Stearic acid is included as a flow agent and silica is employedas flow agent and desiccant.

This example illustrates, as shown in FIG. 3, an increased level ofcontrol possible when release modifying excipients are added to ahydrophilic swellable matrix. The presence of pectin or gelatin isassociated with a degree of pH-dependent degradation and an overallincrease in matrix viscosity which retards the release of the BC. Thisis reflected in the increase in viable CFU delivered after exposure togastric pH. TABLE 3 Dosage Formulas (mg) C1 (CTRL) C2 C3 Lactic acidbacteria pre-blend 150 150 150 HPMC 0 100 100 Pectin 0 100 0 Gelatin 0 0100 Stearic Acid 16 16 16 Silica 16 16 16 TOTAL WEIGHT 182 382 382

EXAMPLE 4

A monolithic tablet of approximately 382 mg containing a hydrophilicagent and a BC was prepared as shown in Table 4 with C1 as the controlgroup. The hydrophilic agent employed is pectin and the lactic acidbacteria pre-blend of lyophilized powder and starch is the BC. Stearicacid is included as a flow agent and silica is employed as flow agentand desiccant.

This example illustrates, as shown in FIG. 4, a level of controlpossible when employing a hydrophilic agent that displays pH-dependentand enzyme-dependent degradation. This example also illustrates the useof a release-modifying agent as a hydrophilic agent. The presence ofpectin is also associated with an overall increase in matrix viscositywhich retards the release of the BC. This is reflected in the increasein viable CFU delivered after exposure to gastric pH. TABLE 4 DosageFormulas (mg) C1 (CTRL) C4 Lactic acid bacteria pre-blend 150 150 Pectin0 200 Stearic Acid 16 16 Silica 16 16 TOTAL WEIGHT 182 382

EXAMPLE 5

A monolithic tablet of approximately 482 mg containing a hydrophilicagent, a release-modifying excipient, an electrolytic agent, and a BCwas prepared as shown in Table 5 with D1 as the control group. Thehydrophilic agent employed is guar gum, the release-modifying excipientemployed is pectin, the electrolytic agent is NaHCO₃, and the lacticacid bacteria pre-blend of lyophilized powder and starch is the BC.Stearic acid is included as a flow agent and silica is employed as flowagent and desiccant.

This example illustrates, as shown in FIG. 5, the application ofgalactomannan gum as a hydrophilic agent in combination with a sodiumsalt and a polysaccharide in a hydrophilic swellable matrix. Thepresence of a galactomannan gum is associated with an overall increasein matrix viscosity which retards the release of the BC, and thepresence of NaHCO₃ is associated with internal pH modulation favorableto the reconstitution of lactic acid bacteria. This is reflected in theincrease in viable lactic acid CFU delivered after exposure to gastricpH. TABLE 5 Dosage Formulas (mg) D1 (CTRL) D2 Lactic acid bacteriapre-blend 150 150 Guar 0 100 NaHCO₃ 0 100 Pectin 0 100 Stearic Acid 1616 Silica 16 16 TOTAL WEIGHT 182 482

EXAMPLE 6

A monolithic tablet of approximately 443 mg containing a hydrophilicagent, an electrolytic agent, a release-modifying excipient, a filler,and a BC was prepared as shown in Table 6. The hydrophilic polymeremployed is HPMC, the electrolytic agent is NaHCO₃, therelease-modifying excipient employed is pectin, the filler employed isMCC and the lactic acid bacteria pre-blend of lyophilized powder andstarch is the BC. The addition of inert filler is associated withincreased power flowability, which is often advantageous duringmanufacture. Stearic acid is included as a flow agent and silica isemployed as flow agent and desiccant. Turmeric is included as acolorant.

As depicted in FIG. 6, the results of this example demonstrate thecapacity for the controlled release of viable BCs over an extendedduration. The controlled release of the hydrophilic matrix is also shownto perform similarly regardless of the duration of exposure to gastricmedia; E1 and E2 are identical formulations showing the difference inrelease based upon a 1 hour, or 2 hour exposure time, respectively.TABLE 6 Dosage Formulas (mg) E1 E2 Lactic acid bacteria pre-blend 150150 HPMC 50 50 NaHCO₃ 50 50 MCC 200 200 Pectin 50 50 Stearic Acid 16 16Silica 16 16 Turmeric 2 2 TOTAL WEIGHT 443 443

EXAMPLE 7

A monolithic tablet of approximately 443 mg containing a hydrophilicagent, an electrolytic agent, a release-modifying excipient, a filler,and a BC was prepared as shown in Table 6.

As depicted in FIG. 7, the results of this example demonstrate thecapacity for the controlled release of bacteria over an extendedduration, for example, from zero to eight hours. The rate of release islinear from zero until approximately 8 hours.

EXAMPLE 8

A monolithic tablet of approximately 532 mg containing a hydrophilicagent, an electrolytic agent, a release-modifying excipient, a filler,and a BC was prepared as shown in Table 8. The hydrophilic agentemployed is HPMC or PEO, the electrolytic agent is NaHCO₃, therelease-modifying excipient employed is pectin, the filler employed isMCC and the bifidobacterium pre-blend of lyophilized powder and starchis the BC. Stearic acid is included as a flow agent and silica isemployed as flow agent and desiccant. Turmeric is included as acolorant.

As depicted in FIG. 8, the results of this example demonstrate thecapacity for the controlled release of BCs over an extended duration.The controlled release of the hydrophilic matrix is also shown torelease in a profile favorable for the delivery of bacteria after eighthours. Such an example would be useful to delivering the bacteria to thelower intestine and beyond. TABLE 8 Dosage Formulas (mg) F2 F3Bifidobacterium bacteria pre-blend 150 150 HIPMC 150 0 PEO 0 150 Pectin100 100 NaHCO₃ 100 100 Stearic Acid 16 16 Silica 16 16 TOTAL WEIGHT 532532

EXAMPLE 9

Two-piece capsules of approximately 665 mg containing two hydrophilicagents, an electrolytic agent, a release-modifying excipient, and a BCwas prepared as shown in Table 9 with G1 as the control group. Thehydrophilic agents employed are HPMC and Guar, the electrolytic agent isNaHCO₃, the release-modifying excipient employed is pectin and thelactic acid bacteria pre-blend of lyophilized powder and starch is theBC. Stearic acid is included as a flow agent and silica is employed asflow agent and desiccant.

This example, as depicted in FIG. 9, demonstrates that the combinationof a hydrophilic agents, an electrolyte, and a release-modifyingexcipient are capable of controlling the release of the BC from acapsule. Dosage form flexibility, such as formulation for a tablet orcapsule, provides substantial adaptability during manufacture. TABLE 9Dosage Formulas (mg) G1 (CTRL) G2 Lactic acid bacteria pre-blend 150 150Pectin 0 75 HPMC 0 110 NaHCO₃ 0 110 Guar 0 200 Stearic Acid 10 10 Silica10 10 TOTAL WEIGHT 170 665

EXAMPLE 10

Monolithic tablets of approximately 684 mg and 342 mg containing ahydrophilic agent, an electrolytic agent, a release-modifying excipient,a filler, and a BC was prepared as shown in Table 10. The hydrophilicpolymer employed is HPMC, the electrolytic agent is NaHCO₃, therelease-modifying excipient employed is pectin, the filler employed isthe MCC, and the lactic acid bacteria pre-blend of lyophilized powderand starch is the BC. Stearic acid is included as a flow agent andsilica is employed as flow agent and desiccant.

This example, as depicted in FIG. 10, demonstrates that the combinationof a hydrophilic agent, and electrolyte, and a release-modifyingexcipient are capable of geometric scalability, tablet shape, size andvolume variation while controlling the release of the BC from thematrix. This flexibility is especially useful in manufacture whendiffering formulation volumes are required when altering tablet shapesand sizes. TABLE 10 Dosage Formulas (mg) H1 H2 Lactic acid bacteriapre-blend 75 150 HPMC 50 100 Pectin 50 100 NaHCO₃ 50 100 MCC 100 200Stearic Acid 8 16 Silica 8 16 Turmeric 1 2 TOTAL WEIGHT 342 684

EXAMPLE 11

Monolithic tablets of approximately 684 mg containing a hydrophilicagent, an electrolytic agent, a release-modifying excipient, a filler,and a BC was prepared as shown in Table 11. The hydrophilic polymeremployed is HPMC, the electrolytic agent is NaHCO₃, therelease-modifying excipient employed is pectin, the filler employed isthe MCC, and the lactic acid bacteria pre-blend of lyophilized powderand starch is the BC. Stearic acid is included as a flow agent and,Silica is employed as flow agent and desiccant. Turmeric is included asa colorant.

This example, as depicted in FIG. 11, demonstrates the application ofdrying an identical formulation of excipients of a pre-blend beforetableting (I2) vs. a non-dried pre-blend (I1). The beneficial effects ofdrying are evidenced by the increase in viable lactic acid bacteria CFUpresent in the dried pre-blend. TABLE 11 Dosage Formulas (mg) I1 I2Lactic acid bacteria pre-blend 150 150 HPMC 100 100 Pectin 100 100NaHCO₃ 100 100 MCC 200 200 Stearic Acid 8 8 Silica 8 8 Turmeric 2 2TOTAL WEIGHT 684 684

EXAMPLE 12

A monolithic tablet of approximately 532 mg containing a hydrophilicagent, an electrolytic agent, a release-modifying excipient, a filler,and a BC was prepared as shown in Table 12. The hydrophilic agentemployed is HPMC of viscosity 4000 mPa or 15000 mPa, the electrolyticagent is NaHCO₃, the release-modifying excipient employed is pectin, thefiller employed is MCC and the bifidobacterium pre-blend of lyophilizedpowder and starch is the BC. Stearic acid is included as a flow agentand silica is employed as flow agent and desiccant. Turmeric is includedas a colorant.

As depicted in FIG. 12, the results of this example demonstrate thecapacity for differential controlled release of viable BCs by employinghydrophilic agents of differing viscosities. TABLE 12 Dosage Formulas(mg) H1 H2 Lactic acid bacteria pre-blend 75 75 HPMC, 4000 mPa 50 0HPMC, 15000 mPa 0 50 Pectin 50 50 NaHCO₃ 50 50 MCC 100 100 Stearic Acid8 8 Silica 8 8 Turmeric 1 1 TOTAL WEIGHT 342 342

EXAMPLE 13

A monolithic tablet of approximately 343 mg containing a hydrophilicagent, an electrolytic agent, a release-modifying excipient, a filler,and a BC was prepared as shown in Table 13. The hydrophilic agentemployed is HPMC, the electrolytic agent is NaHCO₃, therelease-modifying excipient employed is pectin, the filler employed isMCC and the lactic acid pre-blend of lyophilized powder and starch isthe BC. Stearic acid is included as a flow agent and, Silica is employedas flow agent and desiccant. Turmeric is included as a colorant.

As depicted in FIG. 13, the results of this example demonstrate thecapacity for increased stability over time when stored in an ambientenvironment, (25 degrees C., 60% Relative Humidity), evidenced by arelatively constant amount of viable lactic acid bacteria CFU. TABLE 13Dosage Formulas (mg) K1 Lactic acid bacteria pre-blend 75 HPMC 50 Pectin50 NaHCO₃ 50 MCC 100 Stearic Acid 8 Silica 8 Turmeric 2 TOTAL WEIGHT 343

The discussion above is descriptive, illustrative and exemplary and isnot to be taken as limiting the scope defined by any appended claims.

1. A delivery system for a biological component comprising: a deliveryvehicle; and a delivered component, the delivered component including abiologic.
 2. The delivery system of claim 1 wherein the delivery vehicleis a hydrophilic agent, the hydrophilic agent selected from at least oneof the group consisting of a swellable polymer, polysaccharide,polypeptide, resin, and gum.
 3. The delivery system of claim 1 whereinthe delivery vehicle is a hydrophilic agent, the hydrophilic agentselected from at least one of the group consisting of: a) a starchselected from the group consisting of rice, corn or potato starch; b) agum selected from the group consisting of tragacanth gum, locust beamgum, acacia gum, guar gum, xanthan gum, ghatti gum, or galactomannangum; c) an algae derivative selected from alginic acid, sodium alginate,agar, dextran and carageenan; d) a polysaccharide selected from thegroup containing pectin and maltodextrin; e) a cellulose derivativeselected from the group consisting of methylcellulose,carboxymethylcellulose, sodium starch glycollate, sodium or calciumcarboxymethylcellulose, hydroxyethyl methylcellulose, hydroxypropylmethylcellulose, ethylhydroxy ethylcellulose, ethylmethylcellulose,hydroxyethylcellulose, cellulose acetate phthalate or microcrystallinecellulose; f) silica, aluminum silicate, magnesium silicate, aluminummagnesium silicate, sodium silicate or felspar; g) aluminum hydroxide;h) a polypeptide selected from the group consisting of gelatin,collagen, casein or heterogeneous protein mixture; and i) a polymerselected from the group consisting of acrylate, carboxypolymethylene, apolyalkylene glycol or polyvinylpyrrolidone.
 4. The delivery system ofclaim 1 or 2 wherein the delivery system is a monolithic tablet
 5. Thedelivery system of claim 1 or 2 wherein the delivery system is acapsule.
 6. The delivery system of claim 1 or 2 wherein the deliverysystem is an oral delivery system.
 7. The delivery system of claim 1 or2 wherein the biologic is at least a bacteria.
 8. The delivery system ofclaim 1 wherein the delivery vehicle is a hydrophobic agent, thehydrophobic agent selected from at least one of the group consisting ofa wax or other inert material.
 9. The delivery system of claim 1 whereinthe delivery vehicle is a hydrophobic agent, the hydrophobic agentselected from at least one of the group consisting of: a) a wax selectedfrom the group consisting of bees wax and carnuba wax.
 10. The deliverysystem of claim 1 or 8 wherein the delivery vehicle is a monolithictablet
 11. The delivery system of claim 1 or 8 wherein the deliverysystem is an oral delivery system.
 12. The delivery system of claim 1 or8 wherein the biological component includes at least a bacteria.
 13. Adelivery system for a biological component, the system comprising: adelivery vehicle; a release-modifying agent; and a delivered component,the delivered component including a biologic, wherein the deliveryvehicle is a hydrophilic agent.
 14. The delivery system of claim 13wherein the hydrophilic agent is selected from at least one of the groupconsisting of a swellable polymer, polysaccharide, polypeptide, resin,and gum.
 15. The delivery system of claim 13 wherein the hydrophilicagent is selected from at least one of the group consisting of: a) astarch selected from the group consisting of rice, corn or potatostarch; b) a gum selected from the group consisting of tragacanth gum,locust beam gum, acacia gum, guar gum, xanthan gum, ghatti gum, orgalactomannan gum; c) an algae derivative selected from alginic acid,sodium alginate, agar, dextran and carageenan; d) a polysaccharideselected from the group containing pectin and maltodextrin; e) acellulose derivative selected from the group consisting ofmethylcellulose, carboxymethylcellulose, sodium starch glycollate,sodium or calcium carboxymethylcellulose, hydroxyethyl methylcellulose,hydroxypropyl methylcellulose, ethylhydroxy ethylcellulose,ethylmethylcellulose, hydroxyethylcellulose, cellulose acetate phthalateor microcrystalline cellulose; f) silica, aluminum silicate, magnesiumsilicate, aluminum magnesium silicate, sodium silicate or felspar; g)aluminum hydroxide; h) a polypeptide selected from the group consistingof gelatin, collagen, casein or heterogeneous protein mixture; and i) apolymer selected from the group consisting of acrylate,carboxypolymethylene, a polyalkylene glycol or polyvinylpyrrolidone. 16.The delivery system of claim 13 wherein the release-modifying agent isselected from a) an algae derivative selected from alginic acid, sodiumalginate, agar, dextran and carageenan; b) a polysaccharide selectedfrom the group containing pectin and maltodextrin; c) a polypeptideselected from the group consisting of gelatin, collagen, casein orheterogeneous protein mixture; d) a polymer selected from the groupconsisting of acrylate, carboxypolymethylene, a polyalkylene glycol orpolyvinylpyrrolidone; and e) starch selected from the group consistingof rice, corn or potato starch; f) a gum selected from the groupconsisting of tragacanth gum, locust beam gum, acacia gum, guar gum,xanthan gum, ghatti gum, or galactomannan gum; g) a cellulose derivativeselected from the group consisting of methylcellulose,carboxymethylcellulose, sodium starch glycollate, sodium or calciumcarboxymethylcellulose, hydroxyethyl methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, ethylhydroxy ethylcellulose,ethylmethylcellulose, hydroxyethylcellulose, cellulose acetate phthalateor microcrystalline cellulose; h) silica, aluminum silicate, magnesiumsilicate, aluminum magnesium silicate, sodium silicate or felspar; i)aluminum hydroxide; and j) a polymer selected from the group consistingof acrylate, carboxypolymethylene, a polyalkylene glycol orpolyvinylpyrrolidone.
 17. The delivery system of claim 13 wherein thedelivery vehicle is a monolithic tablet
 18. The delivery system of claim13 wherein the delivery vehicle is a capsule.
 19. The delivery system ofclaim 13 wherein the delivery system is an oral delivery system.
 20. Thedelivery system of claim 13 wherein the biologic is at least a bacteria.21. A delivery system for a biological component wherein the systemcomprises: a delivery vehicle; a release-modifying agent; and adelivered component, the delivered component including a biologic,wherein the delivery vehicle is a hydrophobic agent.
 22. The deliverysystem of claim 21 wherein the hydrophobic agent is a wax or other inertmaterial.
 23. The delivery system of claim 21 wherein the hydrophobicagent is a wax, the wax selected from at least one of the groupconsisting of bees wax and carnuba wax.
 24. The delivery system of claim21 wherein the release-modifying agent is a pore-forming excipientselected from at least one of the group consisting of: a) apolysaccharide selected from the group containing pectin andmaltodextrin; b) a cellulose derivative selected from the groupconsisting of methylcellulose, carboxymethylcellulose, sodium starchglycollate, sodium or calcium carboxymethylcellulose, hydroxyethylmethylcellulose, hydroxypropyl methylcellulose, ethylcellulose,ethylhydroxy ethylcellulose, ethylmethylcellulose,hydroxyethylcellulose, cellulose acetate phthalate or microcrystallinecellulose; c) a polymer selected from the group consisting of acrylate,carboxypolymethylene, a polyalkylene glycol or polyvinylpyrrolidone; d)a salt selected from the group consisting of sodium, calcium, potassium,or magnesium salts; e) an algae derivative selected from alginic acid,sodium alginate, agar, dextran and carageenan; and f) starch selectedfrom the group consisting of rice, corn or potato starch.
 25. Thedelivery system of claim 21 wherein the delivery vehicle is a monolithictablet
 26. The delivery system of claim 21 wherein the delivery systemis an oral delivery system.
 27. The delivery system of claim 21 whereinthe biologic includes at least a bacteria.
 28. A delivery system for abiological component wherein the system comprises: a delivery vehicle,the delivery vehicle is a hydrophilic agent; an electrolytic agent; anda delivered component, the delivered component including a biologic. 29.The delivery system of claim 28 wherein the hydrophilic agent isselected from at least one of the group consisting of a swellablepolymer, polysaccharide, polypeptide, resin, and gum.
 30. The deliverysystem of claim 28 wherein the hydrophilic agent is selected from atleast one of the group consisting of: a) a starch selected from thegroup consisting of rice, corn or potato starch; b) a gum selected fromthe group consisting of tragacanth gum, locust beam gum, acacia gum,guar gum, xanthan gum, ghatti gum, or galactomannan gum; c) an algaederivative selected from alginic acid, sodium alginate, agar, dextranand carageenan; d) a polysaccharide selected from the group containingpectin and maltodextrin; e) a cellulose derivative selected from thegroup consisting of methylcellulose, carboxymethylcellulose, sodiumstarch glycollate, sodium or calcium carboxymethylcellulose,hydroxyethyl methylcellulose, hydroxypropyl methylcellulose,ethylhydroxy ethylcellulose, ethylmethylcellulose,hydroxyethylcellulose, cellulose acetate phthalate or microcrystallinecellulose; f) silica, aluminum silicate, magnesium silicate, aluminummagnesium silicate, sodium silicate or felspar; g) aluminum hydroxide;h) a polypeptide selected from the group consisting of gelatin,collagen, casein or heterogeneous protein mixture; and i) a polymerselected from the group consisting of acrylate, carboxypolymethylene, apolyalkylene glycol or polyvinylpyrrolidone.
 31. The delivery system ofclaim 28 wherein the electrolytic agent is selected from at least one ofthe group consisting of a a) a salt selected from the group containingsodium, calcium, potassium, or magnesium salts; b) an amino acid; and c)an ionic compound.
 32. The delivery system of claim 28 wherein thedelivery vehicle is a monolithic tablet
 33. The delivery system of claim28 wherein the delivery vehicle is a capsule.
 34. The delivery system ofclaim 28 wherein the delivery system is an oral delivery system.
 35. Thedelivery system of claim 28 wherein the biological component includes atleast a bacteria.
 36. The delivery system of claim 28 wherein theelectrolytic agent is capable of inducing an intra-dosage form pHphysiologically acceptable to the reconstitution of a lyophilizedbacteria.
 37. A delivery system for a biological component wherein thesystem comprises: a delivery vehicle, the delivery vehicle is ahydrophobic agent; an electrolytic agent; a release modifying agent, anda delivered component, the delivered component including a biologic. 38.The delivery system of claim 37 wherein the hydrophobic agent is a waxor other inert material.
 39. The delivery system of claim 37 wherein thehydrophobic agent is a wax, the wax selected from at least one of thegroup consisting of bees wax, paraffin, and carnuba wax.
 40. Thedelivery system of claim 37 wherein the release-modifying agent is apore-forming excipient selected from at least one of the groupconsisting of: a) a polysaccharide selected from the group containingpectin and maltodextrin; b) a cellulose derivative selected from thegroup consisting of methylcellulose, carboxymethylcellulose, sodiumstarch glycollate, sodium or calcium carboxymethylcellulose,hydroxyethyl methylcellulose, hydroxypropyl methylcellulose,ethylcellulose, ethylhydroxy ethylcellulose, ethylmethylcellulose,hydroxyethylcellulose, cellulose acetate phthalate or microcrystallinecellulose; c) a polymer selected from the group consisting of acrylate,carboxypolymethylene, a polyalkylene glycol or polyvinylpyrrolidone. d)a salt selected from the group consisting of sodium, calcium, potassium,or magnesium salts; e) an algae derivative selected from alginic acid,sodium alginate, agar, dextran and carageenan; and f) starch selectedfrom the group consisting of rice, corn or potato starch.
 41. Thedelivery system of claim 37 wherein the electrolytic agent is selectedfrom at least one of the group consisting of a a) a salt selected fromthe group containing sodium, calcium, potassium, or magnesium salts; b)an amino acid; and c) an ionic compound.
 42. The delivery system ofclaim 37 wherein the delivery vehicle is a monolithic tablet
 43. Thedelivery system of claim 37 wherein the delivery system is an oraldelivery system.
 44. The delivery system of claim 37 wherein thebiologic includes at least a bacteria.
 45. The delivery system of claim37 wherein the electrolytic agent is capable of inducing an intra-dosageform pH physiologically acceptable to the reconstitution of alyophilized bacteria.
 46. An delivery system comprising: a hydrophilicagent; an electrolytic agent; a release-modifying agent; and an activeportion, the active portion including a biological component.
 47. Thedelivery system of claim 46 wherein the hydrophilic agent is selectedfrom at least one of the group consisting of a swellable polymer,polysaccharide, polypeptide, resin, and gum.
 48. The delivery system ofclaim 46 wherein the hydrophilic agent is selected from at least one ofthe group consisting of: a) a starch selected from the group consistingof rice, corn or potato starch; b) a gum selected from the groupconsisting of tragacanth gum, locust beam gum, acacia gum, guar gum,xanthan gum, ghatti gum, or galactomannan gum; c) an algae derivativeselected from alginic acid, sodium alginate, agar, dextran andcarageenan; d) a polysaccharide selected from the group containingpectin and maltodextrin; e) a cellulose derivative selected from thegroup consisting of methylcellulose, carboxymethylcellulose, sodiumstarch glycollate, sodium or calcium carboxymethylcellulose,hydroxyethyl methylcellulose, hydroxypropyl methylcellulose,ethylhydroxy ethylcellulose, ethylmethylcellulose,hydroxyethylcellulose, cellulose acetate phthalate or microcrystallinecellulose; f) silica, aluminum silicate, magnesium silicate, aluminummagnesium silicate, sodium silicate or felspar; g) aluminum hydroxide;h) a polypeptide selected from the group consisting of gelatin,collagen, casein or heterogeneous protein mixture; and i) a polymerselected from the group consisting of acrylate, carboxypolymethylene, apolyalkylene glycol or polyvinylpyrrolidone.
 49. The delivery system ofclaim 46 wherein the release-modifying agent is selected from a) analgae derivative selected from alginic acid, sodium alginate, agar,dextran and carageenan; b) a polysaccharide selected from the groupcontaining pectin and maltodextrin; c) a polypeptide selected from thegroup consisting of gelatin, collagen, casein or heterogeneous proteinmixture; d) a polymer selected from the group consisting of acrylate,carboxypolymethylene, a polyalkylene glycol or polyvinylpyrrolidone; ande) starch selected from the group consisting of rice, corn or potatostarch; f) a gum selected from the group consisting of tragacanth gum,locust beam gum, acacia gum, guar gum, xanthan gum, ghatti gum, orgalactomannan gum; g) a cellulose derivative selected from the groupconsisting of methylcellulose, carboxymethylcellulose, sodium starchglycollate, sodium or calcium carboxymethylcellulose, hydroxyethylmethylcellulose, hydroxypropyl methylcellulose, ethylcellulose,ethylhydroxy ethylcellulose, ethylmethylcellulose,hydroxyethylcellulose, cellulose acetate phthalate or microcrystallinecellulose; h) silica, aluminum silicate, magnesium silicate, aluminummagnesium silicate, sodium silicate or felspar; i) aluminum hydroxide;and j) a polymer selected from the group consisting of acrylate,carboxypolymethylene, a polyalkylene glycol or polyvinylpyrrolidone. 50.The delivery system of claim 46 wherein the electrolytic agent isselected from at least one of the group consisting of a a) a saltselected from the group containing sodium, calcium, potassium, ormagnesium salts; b) an amino acid c) an ionic compound
 51. The deliverysystem of claim 46 wherein the delivery vehicle is a monolithic tablet52. The delivery system of claim 46 wherein the delivery vehicle is acapsule.
 53. The delivery system of claim 46 wherein the delivery systemis an oral delivery system.
 54. The delivery system of claim 46 whereinthe biologic is a bacteria.
 55. The delivery system of claim 46 whereinthe electrolytic agent is capable of inducing an intra-dosage form pHphysiologically acceptable to the reconstitution of a lyophilizedbacteria.
 56. A pre-dosage form blend of the powders, the blendcomprising: about 5% to 40% of an hydrophilic agent by total weight;about 5% to 40% of a release modifying agent by total weight; about 1 to40% of an electrolytic agent by total weight; and a biologicalcomponent.
 57. The pre-dosage blend of claim 56 wherein the pre-dosageblend can be formed into a drug delivery system.
 58. The pre-dosageblend of claim 57 wherein the drug delivery system is monolithicdirectly compressed tablet.
 59. The pre-dosage blend of claim 56 whereinthe hydrophilic agent is at least one of a cellulose derivative andgalactomannan gum.
 60. The pre-dosage blend of claim 59 wherein thecellulose derivative is hydroxypropyl methylcellulose.
 61. Thepre-dosage blend of claim 56 wherein the release-modifying agent atleast one of the group consisting of polysaccharide and a polypeptide.62. The pre-dosage blend of claim 61 wherein the polysaccharide ispectin.
 63. The pre-dosage blend of claim 56 wherein the electrolyticagent is selected from at least one of the group consisting of sodiumcarbonate, sodium biocarbonate, sodium phosphate, and calcium carbonate.64. The pre-dosage blend of claim 56 wherein the biologic is aprobiotic.
 65. The pre-dosage blend of claim 64 wherein probiotic abacteria.
 66. A the method of making an extended release dosagecomprising: desiccating at least one of the group consisting of arelease modifying agent, electrolyte, and a hydrophilic agent; addingthe desiccated material to a biologic, whereby a reduction in theavailable water content of the dosage is produced.